Division of Molecular Embryology, DKFZ-ZMBH Alliance, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Dev Dyn. 2011 Jan;240(1):188-94. doi: 10.1002/dvdy.22500.
Non-canonical/planar cell polarity (PCP) Wnt signaling plays important roles in embryonic development and tissue homeostasis, and is implicated in human disease. Monitoring Wnt/PCP signaling relies mostly on semi-quantitative bioassays or biochemical analysis. Here we describe a luciferase reporter assay based on an ATF2 response element, which faithfully monitors non-canonical Wnt signaling in Xenopus embryos. The assay is simple, quantitative, and robust. It can be used to detect non-canonical Wnt signaling changes following gain and loss of function of pathway components, including Wnt, Frizzled, Ror2, Disheveled, Rac1, MKK7, and JNK. Wnt/PCP signaling has recently been implicated in left-right asymmetry and our reporter assay suggests that in gastrula embryos there is a right-ward bias in Wnt/PCP signaling. We also mapped Wnt/PCP signaling in the early Xenopus embryo and find that it peaks in the dorso-vegetal region, paralleling Wnt/β-catenin signaling.
非经典/平面细胞极性（PCP）Wnt 信号在胚胎发育和组织稳态中发挥重要作用，并与人类疾病有关。Wnt/PCP 信号的监测主要依赖于半定量生物测定或生化分析。在这里，我们描述了一种基于 ATF2 反应元件的荧光素酶报告基因检测，该检测能在非洲爪蟾胚胎中忠实地监测非经典 Wnt 信号。该检测方法简单、定量且稳健。它可用于检测通路成分（包括 Wnt、Frizzled、Ror2、Disheveled、Rac1、MKK7 和 JNK）获得或功能丧失后非经典 Wnt 信号的变化。Wnt/PCP 信号最近与左右不对称有关，我们的报告基因检测表明，在原肠胚中，Wnt/PCP 信号存在向右偏倚。我们还在早期非洲爪蟾胚胎中对 Wnt/PCP 信号进行了定位，发现它在背-腹区域达到峰值，与 Wnt/β-catenin 信号平行。
